Anti-MIRO1 antibody [MJF-D29136-4] - BSA and Azide free

• I-ELISA

Supplier Data

Indirect ELISA - Anti-MIRO1 antibody [MJF-D29136-4] - BSA and Azide free (AB319155)

This data was developed using ab319154, the same antibody clone in a different buffer formulation.

Indirect ELISA analysis of ab319154 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Mouse RHOT1.

Antigen concentration : 1000 ng/ml

• WB

Supplier Data

Western blot - Anti-MIRO1 antibody [MJF-D29136-4] - BSA and Azide free (AB319155)

This data was developed using ab319154, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the higher MW band at approximately 150 kDa (in lane 2) is unknown.

All lanes:

Western blot - Anti-MIRO1 antibody [MJF-D29136-4] (<a href='/en-us/products/primary-antibodies/miro1-antibody-mjf-d29136-4-ab319154'>ab319154</a>) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Lane 3:

Human cerebellum tissue lysate at 20 µg

Lane 4:

Mouse cerebellum tissue lysate at 20 µg

Lane 5:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 6:

Mouse lung tissue lysate at 20 µg

Lane 7:

Rat cerebellum tissue lysate at 20 µg

Lane 8:

Rat skeletal muscle tissue lysate at 20 µg

Lane 9:

Rat lung tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG

Observed band size: 80 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-MIRO1 antibody [MJF-D29136-4] - BSA and Azide free (AB319155)

This data was developed using ab319154, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-MIRO1 antibody [MJF-D29136-4] (<a href='/en-us/products/primary-antibodies/miro1-antibody-mjf-d29136-4-ab319154'>ab319154</a>) at 1/1000 dilution

Lane 1:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

mIMCD3 (mouse inner medullary collecting duct epithelial cell) whole cell lysate at 20 µg

Lane 3:

4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 6:

MEF (mouse embryo fibroblast) whole cell lysate at 20 µg

Lane 7:

A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate at 20 µg

Lane 8:

L6 (rat skeletal muscle myoblast) whole cell lysate at 20 µg

Lane 9:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 80 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-MIRO1 antibody [MJF-D29136-4] - BSA and Azide free (AB319155)

This data was developed using ab319154, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab319154 was shown to bind specifically to RHOT1. Target of interest was observed at 80 kDa band in wild-type Hela cell lysates (lane 1) with no signal observed at this size in RHOT1 knockout cell line (lane 2) (lane 2, knockout cell line ab265792).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-MIRO1 antibody [MJF-D29136-4] (<a href='/en-us/products/primary-antibodies/miro1-antibody-mjf-d29136-4-ab319154'>ab319154</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human RHOT1 (MIRO1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-rhot1-miro1-knockout-hela-cell-line-ab265792'>ab265792</a>) at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 80 kDa,36 kDa

false

Exposure time: 48s

• I-ELISA

Supplier Data

Indirect ELISA - Anti-MIRO1 antibody [MJF-D29136-4] - BSA and Azide free (AB319155)

This data was developed using ab319154, the same antibody clone in a different buffer formulation.

Indirect ELISA analysis of ab319154 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution.

Antigen : Human RHOT1,Human RHOT2.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with human RHOT2.